pPPEM
(Plasmid #162467)

• Purpose: Destination Expression vector for monocot plant primer editing
• Depositing Lab: Adam Bogdanove
• Publication: Wang et al Front Genome Ed. 2021 Mar 4;3:617553. doi: 10.3389/fgeed.2021.617553. eCollection 2021.

Backbone

• Vector backbone: pCAMBIA1300
• Backbone size w/o insert (bp): 8959
• Total vector size (bp): 18478
• Vector type: Plant Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Cas9H840A
• Species: Synthetic
• Insert Size (bp): 10576
• Mutation: Change histidine 840 to Alanine
• Promoter: maize Ubi
• Tag / Fusion Protein:
  • MLV reverse transcriptase (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Acc65I (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: ATCCGATCCAAATCCGAA
• 3′ sequencing primer: TTCGAATTCCGAACCGTC

Resource Information

• Supplemental Documents:
  • pPPEM.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The original vector is from Dr. Bing Yang at Iowa State University. We modified it. Reference:
Large chromosomal deletions and heritable small genetic changes induced by CRISPR/Cas9 in rice. Zhou, H., Liu, B., Weeks, D. P., Spalding, M. H., Yang, B. Nucleic Acids Res. 2014, 42(17):10903-14. Please visit https://www.biorxiv.org/content/10.1101/2020.07.16.206276v1 for bioRxiv preprint.